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SLIC: A Method for Sequence- and Ligation-Independent Cloning

  • Mamie Z. Li
  • Stephen J. Elledge
Part of the Methods in Molecular Biology book series (MIMB, volume 852)

Abstract

We describe here a method for sequence- and ligation-independent cloning (SLIC). SLIC uses an exonuclease, T4 DNA polymerase, to generate single-stranded DNA overhangs in insert and vector sequences. These fragments are then assembled in vitro and transformed into Escherichia coli to generate recombinant DNA of interest. SLIC inserts can also be generated by incomplete PCR (iPCR) or mixed PCR. As many as five inserts can be assembled in one reaction simultaneously with great efficiency using SLIC. SLIC circumvents sequence constraints for recombinant DNA using standard restriction enzyme-mediated cloning and previous ligation-independent cloning methods and provides a new approach for the efficient generation of recombinant DNA.

Key words

SLIC Ligation-independent cloning In vitro homologous recombination iPCR Sequence-independent cloning Subcloning 

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Copyright information

© Springer Sceince+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of GeneticsHoward Hughes Medical Institute, Harvard Medical SchoolBostonUSA
  2. 2.Division of Genetics, Department of MedicineBrigham and Women’s HospitalBostonUSA

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